Journal:

Article Title: A protocol for a lung neovascularization model in rodents

doi: 10.1038/nprot.2007.536

Figure Lengend Snippet: Drawing illustrating the inflation apparatus to distend lungs initially with a 3% paraformaldehyde/0.1% gluteraldehyde solution (shown in blue). Fixative is delivered to the tracheal catheter at 23 cm H2Opressure (determined by gravity) and to the vascular catheter at 100 cm H2O (determined by positive pressure from a peristaltic pump). Note: use a standard pressure monometer to predetermine the height of the mercury column needed for the pump to generate this pressure. The catheters connect first to a luer adapter, or a luer-adapted needle, and then, via a 4-way stopcock, to the tubing delivering fixative from each reservoir. Hemostat clamps (indicated by black bars) regulate the flow of fixative through the delivery tubes from each reservoir. A gate clip (heavy black bar) positioned on the tubing from the peristaltic pump is used to obtain/regulate pressure in the vascular reservoir.

Article Snippet: 0.28 mm) Luer adapter needles (Beckton-Dickinson for rat trachea and pulmonary artery—16 gauge stub adapter, cat. no. 427561; for mouse trachea, 22G, 0.7 mm × 25.4 mm, cat. no. 305155; and for pulmonary artery, 27G, 0.4 mm × 12.7 mm, cat. no. 305109) 3-way stopcock with male luer-lock adapter (Mallinckrodt, cat. no. 91040) Sterile specimen containers, 4 oz (Fisher Scientific, cat. no. 14-375-147) Single-edged industrial blades (Ted Pella, cat. no. 121-72) Cotton gauze sponges, 2 inch × 2 inch (Fisher Scientific, cat. no. 22-362-178) Kendall Tendersorb underpads, 17 inch × 24 inch (Owens & Minor, cat. no. 3583 007105) Single-use disposable scalpels, no. 15 blade (Fisher Scientific, cat. no. 08-927-5C) Shell vials, 15 mm o.d.

Techniques:

Journal: Nature Communications

Article Title: Pancreatic tumor eradication via selective Pin1 inhibition in cancer-associated fibroblasts and T lymphocytes engagement

doi: 10.1038/s41467-022-31928-7

Figure Lengend Snippet: a Establishment and validation of CAFs via high expressions of both FAP-α and α-SMA. b Cellular uptake of Cy5-labeled formulations on Pan02 cells, NIH-3T3 cells, and CAFs, measured by flow cytometry ( n = 3 biologically independent experiments; error bars, mean ± SD). c Cellular uptake of Cy5-labeled antiCAFs-DMS on cells, imaged by fluorescent microscopy. Scale bars are 20 μm. Three independent repetitions got similar results. d Cellular uptake of Cy5-labeled antiCAFs-DMS on cells, quantified by flow cytometry ( n = 6 biologically independent samples; error bars, mean ± SD). Cytotoxicity on Pan02 cells ( e ) and CAFs ( f ) ( n = 3 biologically independent experiments; error bars, mean ± SD). g Western blotting detection of Pin1 and relative proteins from Pan02 cells after treating with PBS, AG17724 (0.5 μM), or antiCAFs-DMS (corresponds to 0.5 μM of AG17724) for 24 hours. h Quantitative analysis of the western blotting results from Pan02 cells (Three independent repetitions got similar results.). i Western blotting detection of Pin1 and relative proteins from CAFs treated with PBS, AG17724, or antiCAFs-DMS after treating with PBS, AG17724 (0.5 μM) or antiCAFs-DMS (corresponds to 0.5 μM of AG17724) for 24 hours. j Quantitative analysis of the western blotting results from CAFs (Three independent repetitions got similar results.). One-way analysis of variance followed by Turkey posttests. Source data are provided as a Source Data file.

Article Snippet: Anti-FAP-α antibody (MAB9727-100, 0.25 μg/mL as working concentration.) and anti-phospho-CDC2/CDK1 (Y15) antibody (AF888-SP, 0.2 μg/mL as working concentration.) were purchased from R&D SYSTEMS.

Techniques: Biomarker Discovery, Labeling, Flow Cytometry, Microscopy, Western Blot